This invention relates to a method for predicting the constitution susceptible to the onset of specific diseases, effects on methods of treatment for patients suffering from said diseases or predicting the prognosis of the treatment by analysis of genetic polymorphisms of a human trypsin-like enzyme of a respiratory tract.
Research on related genes has recently been promoted not only in genetic diseases due to deletion or mutation of single genes but also in multifactorial diseases caused by entanglement of several genetic predispositions and environmental factors. As a result, the deletion or point mutation and isoforms related to the multifactorial diseases and further mutation of genetic parts (introns or promoters) without affecting actually translated amino acid sequences have come to be considered as risk factors for the diseases.
It has been published that the correlation is recognized between bone density and genetic polymorphisms of an intron of the vitamin D receptor in the osteopathic field as the prior art (Morrison, N. A. et al., Nature, 367: 284-287 1994). In the field of circulatory organs, it has been reported that the I type (insertion type) and D type (deletion type) genetic polymorphisms of an angiotensin-converting enzyme are associated with the onset of myocardial infarctions (Cambien, F. et al., Nature 359: 641-644, 1992) and the amino acid substitution of M295T of angiotensinogen and the polymorphisms of a promoter region of G-6A are associated with the onset of essential hypertension (Inoue, I. et al., J. Clin. Invest., 99: 1786-1797, 1997). Furthermore, in the field of the nervous system, it has been reported on the association between the onset of dementia and the isoforms of apoE protein. Much research has been carried out in the association between the genetic polymorphisms of glutathione S-transferase and the onset of cancers in the cancer-related field. As the field of respiratory diseases, it has been reported on the association between the onset or morbid state of asthma and the TNF (Moffatt, M. F. et al., Hum. Mol. Genet. 6 (4): 551-554, 1997) and the association between the onset or morbid state of the asthma and the genetic polymorphisms of an angiotensin converting enzyme (Benessiano, J. et al., J. Allergy Clin. Immunol. 99 (1): 53-57, 1997).
Furthermore, attention has been paid to the genetic background as one of causes of difference between patients in sensitivity to drugs used for treating diseases, and it is has been desired even by the medical site to provide the directionality such as selection of methods of treatment according to the drug sensitivity of individual humans by diagnosis of the genetic polymorphisms. It is thought that the drug development by selecting patient groups expectable of drug effects according to the genetic polymorphisms is effective in clinical trials (Kleyn K. W. et al., Science, 281: 1820-1821, 1998). A report on the genetic polymorphisms of an intron of an angiotensin converting enzyme [ACE (angiotensin converting enzyme)] and effects of an ACE inhibitor (Yoshida, H. et al., J. Clin. Invest. 96: 2162-2169, 1995), a report on the genetic polymorphisms of beta 2-adrenergic receptor and effects of the beta-agonist on asthma (Liggett, S. B., Am. J. Respir. Crit. Care Med. 156 (4 Pt 2): S156-162, 1997) and the like are cited as the conventional reports related to the drug sensitivity and the genetic polymorphisms.
On the other hand, the human trypsin-like enzyme of the respiratory tract related to the present invention has been purified from the sputum of patients suffering from chronic airway diseases (Japanese Unexamined Patent Publication No. 7-067640 and Yasuoka, S. et al., Am. J. Respir.Cell Mol. Biol., 16: 300-308, 1997) and the amino acid sequence and cDNA sequence thereof have been already made clear (Japanese Unexamined Patent Publication No. 8-89246 and Yamaoka K. et. al., J. Biol. Chem., 273(19): 11895-11901, 1998). Several studies have been made of the activity possessed by the enzyme in vitro. Since the enzyme has production enhancing actions on cytokines such as IL-8 or GM-CSF derived from a human bronchial epithelial cell line including the association with mucociliary movement, the possibility for association with the morbid state of airway inflammations is considered (Terao, Noriko et al., the Japanese Respiratory Society, 1998). Since the enzyme has enzyme activities such as hydrolytic activity for fibrinogen and activating actions on plasminogen activators (pro-urokinase) (Yoshinaga, Junko et al., Conference on Proteases and Inhibitors in Pathophysiology and Therapeutics, 1998), the possibility is assumed for anti-inflammatory actions through the formation of fibrins on the airway mucosal surfaces or modification of the morbid state thereof in chronic airway diseases and the possibility is also considered for the association with cancer metastasis or the like. The association of the enzyme with physiological functions or the morbid state in vivo is not yet sufficiently elucidated, and the genetic parts (introns or promoters) without corresponding to the actually translated amino acid sequences has not yet known about genes at all. Further, no investigation has hitherto been carried out on the association of the presence or absence of the genetic polymorphisms for the human trypsin-like enzyme of the respiratory tract or the genetic polymorphisms with diseases.
By the way, much information can be provided about the prediction of the onset of specific diseases, prognosis of treatment, selection of appropriate methods of the treatment and administered drugs or the like by the prediction of diseases-associated constitution by genetic analysis. Accordingly, the prediction is desired by many physicians and patients and further makes the prophylaxis of onset and early therapy possible. Therefore, it is thought that the prediction is related with a reduction in medical care expenditures to become indispensable for the future medical care.
It is, however, very difficult to find out a gene associated with diseases and establish an analytical method therefor, and there are few examples of genetic analytical methods in which the association with diseases is recognized as described above. Therefore, the development of the genetic analytical technique for making various disease-associated constitutions predictable is strongly desired.
On the other hand, it is not yet elucidated with what diseases the human trypsin-like enzyme of the respiratory tract is associated at present.
As a result of intensive research made in consideration of the problems of the prior art, the present inventors et al. have designed a primer for genetically amplifying an intron part on the genome of the human trypsin-like enzyme of the respiratory tract specifically expressing in the human respiratory tract, novelly determined the DNA sequence of both the termini of the amplified genetic fragment and the novelly determined DNA sequence in an exonlintron border part, found out that there are genetic polymorphisms in the amplified genetic fragment and the novelly determined DNA sequence and further firstly found out the association of the genotypes of the human trypsin-like enzyme of the respiratory tract with diseases in individual humans by analyzing the genetic polymorphisms, thus attaining the present invention.
That is, an object of the present invention is to provide a method for predicting the constitution of individual humans susceptible to the onset of specific diseases or effects of treatment on patients or prognosis of the treatment by analyzing the genetic polymorphisms of the human trypsin-like enzyme of the respiratory tract.
Further, the present invention is a method for diagnosing an abnormality in the mucociliary biophylactic system by the analysis of the genetic polymorphisms of the human trypsin-like enzyme of the respiratory tract and a method for predicting the constitution of individual humans susceptible to the onset of diseases, effects of treatment on patients or prognosis of the treatment.
Furthermore, the present invention is a genetic fragment containing a part or all of the base sequence of an intron in the human trypsin-like enzyme of the respiratory tract.